Use of coisogenic host blastocysts for efficient establishment of germline chimeras with C57BL/6J ES cell lines
Use of coisogenic host blastocysts for efficient establishment of germline chimeras with C57BL/6J ES cell lines
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DOI：
10.2144/01315st01
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发表时间：
2001-11-01
期刊：
影响因子：
2.7
通讯作者：
Conover, JC
中科院分区：
文献类型：
--
作者：
Schuster-Gossler, K;Lee, AW;Conover, JC
在胚胎干细胞中进行基因打靶，可以产生具有特定基因突变的小鼠。目前，只有有限数量的小鼠品系才能获得具有生殖系能力的ES细胞系，而不适当的ES细胞/宿主囊胚组合往往限制了基因靶向小鼠的高效生产。在这里，我们描述了C57BL/6J(B6)ES系的建立，并比较了两个宿主囊胚供体FVB/NJ(FVB)和同基因菌株C57BL/6-Tyr(c-2J)(c(2j))在生产生殖系嵌合体方面的效果。我们发现，将B6 ES细胞注射到C(2J)宿主囊胚中，可以检测到很高的毛色嵌合率，并且可以在很少的囊胚注射下获得生殖系传递。在所有的病例中，除了一例外，高度嵌合的小鼠都100%地传播给了它们的后代。将B6 ES细胞注射到FVB囊胚中，产生了一些嵌合小鼠。然而，毛色嵌合体的比例很低，需要更多的胚泡注射才能产生能够通过生殖系传播的嵌合体。我们的数据支持使用同基因白化宿主菌株c(2J)在使用B6 ES细胞时产生具有生殖系能力的嵌合小鼠。
Gene targeting in embryonic stem (ES) cells allows the production of mice with specified genetic mutations. Currently, germline-competent ES cell lines are available from only a limited number of mouse strains, and inappropriate ES cell/host blastocyst combinations often restrict the efficient production of gene-targeted mice. Here, we describe the derivation of C57BL/6J (B6) ES lines and compare the effectiveness of two host blastocyst donors, FVB/NJ (FVB) and the coisogenic strain C57BL/6-Tyr(c-2J) (c(2J)), for the production of germline chimeras. We found that when B6 ES cells were injected into c(2J) host blastocysts, a high rate of coat-color chimerism was detected, and germline transmission could be obtained with few blastocyst injections. In all but one case, highly chimeric mice transmitted to 100% of their offspring. The injection of B6 ES cells into FVB blastocysts produced some chimeric mice. However, the proportion of coat-color chimerism was low, with many more blastocyst injections required to generate chimeras capable of germline transmission. Our data support the use of the coisogenic albino host strain, c(2J), for the generation of germline-competent chimeric mice when using B6 ES cells.